Treatment of hyperlipidaemia

ABSTRACT

3-Tert.-butyl-4-hydroxyacetanilides are administered to mammal, especially human, subjects to reduce serum lipid, especially cholesterol, levels. 3,5-Di-tert.-butyl-4-hydroxyacetanilide is preferred and the invention has particular application to the treatment of hypercholesterolaemia in humans.

Corne United States Patent [191 TREATMENT OF HYPERLIPIDAEWA [75]Inventor: Seymour Jeffrey Corne, Kew,

England [73] Assignee: Aspro-Nicholas Limited, Slough,

England 22 Filed: Mar. 1, 1974 [21] Appl. No.: 447,453

[30] Foreign Application Priority Data Mar. 7, 1973 United Kingdom11070/73 Mar. 7, 1973 United Kingdom ll07l/73 [52] US. Cl. 424/324 [51]Int. Cl. A61K 31/165 [58] Field of Search 424/324 [56] References CitedUNITED STATES PATENTS 3,492,349 I/ 1970 Doyle et a1. 424/324 FOREIGNPATENTS OR APPLICATIONS 1,208,710 10/1970 United Kingdom PrimaryExaminer-Donald B. Meyer [57] ABSTRACT 8 Claims, No Drawings 1 TREATMENTOF HYPERLIPIDAEMIA The present invention relates to the reduction ofserum lipid levels in animals and in particular provides a treatment forhyperlipidaemia, especially hypercholesterolaemia, i.e., excess. ofcholesterol in the blood.

UK. Pat. Specification No. 1208710 (Aspro- Nicholas Limited),publishedon Oct. 14, 1970, discloses that certain3,5-dialkyl4-hydroxyacetanilides, especially the 3,5-ditert,-butylcompound, possess a higher level of analgesic activity than paracetamol(i.e., 4-hydroxyacetanilide) and/or more slowly excreted thanparacetamol. It has now been found that 3,5-ditert.butyl-4-hydroxyacetanilide and other 3-tert.-butyl'4-hydroxyacetanilides optionally substituted in the aromaticnucleus by C 1 to C alkyl groups have antihyperlipidaemic activity asdetermined in a screening test hereinafter described.

-According to the present invention therefore, there is provided amethod of reducing serum lipid levels in a mammal, especially thosehaving hyperlipidaemia and more especially hypercholesterolaemia, whichcomprises administering to the animal subject a lipidreducing amount ofa 3-tert-butyl-4-hydroxyacetanilide of the formula 1:

C(CH

NHCOCH3 wherein R represents a hydrogen atom or an alkyl groupcontaining one to four carbon atoms.

The alkyl group represented by R may be any straight or branched chainalkyl group containing from one to four carbon atoms and may be locatedin any one of the 2, 5 and 6 positions of the aromatic nucleus,preferably the 5 or 6 position. Alkyl groups having from one to fourcarbon atoms are methyl, ethyl, nand iso-propyl, and n-, sec.- andtert.-butyl.

The presently preferred compound of formula 1 is3,5-di-tert.-butyl-4-hydroxyacetanilide. Other compounds of formula 1include 3-tert.-butyl-6-methyl-4 hydroxyacetanilide3-tert.-butyl-4-hydroxyacetanilide 3-tert.-butyl-5-sec.-propyl-4-hydroxyacetanilide 3-tert.-butyl-6-ethyl-4-hydroxyacetanilide3-tert.butyl-2-methyl-4-hydroxyacetanilide3,6-ditert.-butyl-4-hydroxyacetanilide The alkyl group R and thearomatic nucleus of the compounds of formula 1 may be substituted withone or more substituents which are therapeutically compatible with themolecule being substituted.

The term therapeutically compatible as used in this specification inrelation to a substituent means that the presence of that substituentneither destroys the pharmacological activity of the molecule nor sodecreases said activity and/or increases the toxicity of the moleculethat the therapeutic ratio is reduced .to five or below. The therapeuticcompatibility of a particular substituent maydepend upon the intendedsite bf substitution in the molecule and/or the presence in the moleculeof other substituents. Hence a given substituent may be therapeuticallycompatible in respect of one molecule into which it is to be introducedbut incompatible, i.e., inactivating, in respect of another molecule.The compatibility of any substituent in respect of any molecule havingthe basic formula 1 can be readily assessed by subjecting the relevantcompound to standard screening tests such as that referred tohereinafter. It is well-within the ability of the averagely skilled manconcerned withthe development of new drugs to ascertain whichsubstituents may be present and at what positions in pharmaceuticallyactive compounds of formula 1.

The 3-tert.-butyl-4-hydroxyacetanilides of formula 1 can be prepared bythe methods described in U.K. Pat. Specification Nos. 1198059(AsproNicholas Limited) and 1208710 (Aspro-Nicholas Limited) from thecorresponding 4-hydroxyanilines.

The effect of the 3-tert.-butyl-4-hydroxyacetanilides of formula 1(hereinafter referred as the active compounds of the invention) on serumlipid levels and in particular their anti-hypercholesterolaemic activity(hereinafter referred to as AHCA) has been assessed on the basis ofantagonism of Triton WR 1339-induced hypercholesterolaemia in the rat.

Triton WR 1339 is an oxyethylated tertiary octyl'phenol polymethylenepolymer supplied by Ruger Chemical Co. Inc, It has been found that ifrats" which have been fasted overnight (about 16 hours) are injectedintravenously with Triton WR 1339 in an amount of 200 mg/kg body weight,the serum cholesterol levels after 6 hours are some 2 to 3 times thoseof untreated control rats. Certain drugs, with the notable exception ofAtromid-S (i.e., clofibrate), which are known to reduce serumcholesterol levels in humans antagonise said Triton-inducedhypercholesterolaemia.

In the screening test, used to assess AHCA the test compound wassuspended in 0.5% w/w aqueous gum tragacanth and then administeredorally in an amount of 1 ml. suspension per g. body weight to"a firstgroup of six male albino rats which had been fasted overnight. One hourlater each of the rats was injected via its tail vein with 0.2 ml per100 g body weight of a 10% w/v solution of Triton WR 1339; the ratshaving first been warmed to about 30C for 15 minutes. After the lapse of6 hours from injection, the rats were bled by direct heart punctureunder either anaesthetic and serum cholesterol levels measured using themethod of Zlatkis et al (Journal of Laboratory and Clinical Medicine,41, 1953, 486). Each of a second group of six male albino rats wasinjected with Triton WR 1339 as above but without the prior oraladministration of the test compound.

A third group of six male albino rats was used as a control r In eachgroup, the rats wereof the Carworth Europe CFHB strain and of a weightin excess of 300 g.

The mean value of serum cholesterol level in each group was determinedfrom a standard curve constructed from the individual results of thegroup and the percentage inhibition was calculated according to theformula.

where C, represents the mean value for the first group;

C represents the mean value for the second group;

and

C represents the mean value for the third group.

When a typical active compound of formula 1, viz.3-tert.-buty-l-6-methyl-4-hydroxyacetanilide, was submitted to theaforementioned screening test, the percentage inhibition at a dose levelof lOOmg/kg was calculated to be 19.3. On repeated submission of thepreferred L compound, viz. 3,5-di-te1t.butyl-4- hydroxyacetanilide tosaid screening test, the percentage inhibition at a dose level of 100mg/kg was found to be in the range 25.8 to 33.4. These results comparewith a percentage inhibition at the same dose level of 34 calculatedfrom a comparative test with 3-methylsalicylic acid. The latter compoundwas known to have a high AHCA from the work of Hyams and Howard (ModernGeriatrics, July 1971, 336/342).

Another comparative test was carried out using 100 mg/kg of3,5-di-iso-propyl-4-hydroxyacetanilide as the test compound. Thiscompound was selected because of its close structural resemblance to theactive compound of the present invention and, as far as we are aware,its AHCA had not previously been assessed. In this test, no antagonismof the Triton-induced antihypercholesterolaemia occurred.

The 3ten.-butyl-4-hydroxyacetanilides of formula 1 are usually insolublein most pharmaceutically acceptable solvents and hence usually will beadministered rectally or, more usually, orally; Generally they will beadministered in association with a pharmaceutically acceptable carrier.For example, they can be mixed with a carrier, suspended in a carrier orenclosed or encapsulated by a carrier in the form of a capsule, sachet,cachet, paper or other container. The carrier may be a solid, semi-solidor liquid material which serves as a vehicle, excipient or diluent forthe active material. Some examples of the carriers which may be employedare lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia,calcium phosphate, liquid paraffin, cocoa butter, oil of theobroma,alginates, gum tragacanth, gelatin, Syrup B.P., methyl cellulose,polyoxyethylene sorbitan monolaurate, and methyl and propylhydroxybenzoates. In particular the formulations containing the activecompounds of the present invention may be in the form of tablets,capsules and suspensions.

The dosage range within which a therapeutically useful reductionin'serum lipid levels is produced by administration of the3-tert.-butyl-4-hydroxyacetanilides of formula 1 will vary according tointer alia the species and weight of the animal to be treated and theroute of administration as is well known in the art. Such a range willusually fall within the limits of 0.01 to 250 mg/kg. A preferred rangefor rats is 100 to 200 mg/kg. In the case of humans it is expected thatsaid dosage for adult humans will be within the range of l to 4 g perday.

The following Examples illustrate typical formulations in which theactive compound of the present invention can be administered to humans.All parts are by weight unless otherwise stated.

EXAMPLE 1 Tablets containihg the following components3,5-di-tert.-butyl-4-hydroxy-acetanilide 1000 parts Starch 1 parts Talc5 parts Magnesium stearate 5 parts a 8.8. No. 40 sieve, the granulesmixed with the remainder of the starch, and the talc and magnesiumstearate added. The resultant mixture is compressed into tabletsweighing (a) 110 mg, (b) 275 mg., and (c) 550 mg, to provide tabletscontaining respectively 100, 250 and 500 mg of3,5-di-tert.-butyl-4-hydroxyacetanilide.

Two of the 500 mg tablets administered three times daily will reduce thecholesterol level in patients suffering from hypercholesterolaemia.Other dosage regimes of between 1 to 4 g daily may readily be maintainedusing the 100, 250 and 500 mg tablets separately or in combination.Control of blood lipid levels is usually necessary over a prolonged ifnot indefinite period and hence an appropriate dosage regime may beestablished for the individual patient.

EXAMPLE 2 Capsules each containing the following components are made3,5-di-tert.-butyl-4-hydroxyacetanilide parts Calcium phosphate 20 partsThe powders are thoroughly mixed together and filled into hard gelatincapsules so that each capsule contains 250 mg of 3,5-di-tert.-butyl4-hydroxyacetanilide.

EXAMPLE 3 Suppositories each having the following composition are madeup as follows:

10 parts 30 parts 2,5-di-tert.-4-hydroxyacetanilide Oil of Theobroma Thefinely powdered 3,5-di-tert.-butyl-4- hydroxyacetanilide is trituratedwith the molten oil of theobroma and the mixture so produced poured intosuppository moulds of a nominal capacity of l g or 2 g as desired toproduce suppositories each containing re spectively 250 mg or 500 mg of3,5-di-tert.-butyl-4- hydroxyacetanilide.

The caps'ulesand suppositories of Examples 2 and 3 may be used insteadof the tablets of Example 1 to maintain the dosage regimes referred toin that Example. Having regard to the long term treatment required formost patients, the oral forms of Examples 1 and 2 are preferred to therectal form of Example 3'.

Having regard to the foregoing disclosure, thefollow- 7 ing is claimedas the inventive and patentable embodiments thereof:

l. A method of reducing serum cholesterol in a mamwherein R is in the or6 position and represents a hydrogen atom or an alkyl group of one tofour carbon atoms.

2. The method according to claim 1 wherein the 3-tert.-butyl-4-hydroxyacetanilide is 3,5-di-tert.-butyl-4-hydroxyacetanilide.

3. The method according to claim 1 wherein the 3-tert.-butyl-4-hydroxyacetanilide is 3-tert.-butyl-6- hydroxyacetanilide.

CERTIFICATE OF CORRECTION Paten: 310. 3, 899, 596 D t d August 12, I975Inve:cor(s) Seymour Jeffrey Corne Q It is certified that error appearsin the above-identified patent and that said Letters Patent are herebycorrected as shown below:

Column 1, line 13 after "and/or" please insert --are-- W C Column 2,line 68 7r Inhibition X00 should be e v (C C2) Inhibition 100 1 a Signedand 5231211 this I twentieth Day Of April 1976 k [SEAL] Arrest:

RUTH C. MASON C. MARSHALL DANN Alusn'ng Officer (mnml'xsl'unvrnj'lau'nls and Trademarks i

1. A METHOD OF REDUCING SERUM CHOLESTEROL IN A MAMMAL WHICH COMPRISESADMINISTERING TO THE MAMMAL IN NEED THEREOF A CHOLESTEROL-REDUCINGEFFECTIVE AMOUNT OF A 3-TERT-BUTYL-4HYDROXYACETANILIDE OF THE FORMULA:2. The method according to claim 1 wherein the3-tert.-butyl-4-hydroxyacetanilide is3,5-di-tert.-butyl-4-hydroxyacetanilide.
 3. The method according toclaim 1 wherein the 3-tert.-butyl-4-hydroxyacetanilide is3-tert.-butyl-6-methyl-4-hydroxyacetanilide.
 4. The method according toclaim 1 wherein the 3-tert.-butyl-4-hydroxyacetanilide is administeredorally.
 5. The method according to claim 1 wherein the3-tert.-butyl-4-hydroxyacetanilide is administered rectally.
 6. Themethod according to claim 1 wherein the mammal is human havinghypercholesterolaemia.
 7. The method according to claim 6 wherein the3-tert.-butyl-4-hydroxy acetanilide is administered in a daily amount offrom 1 to 4 g via doses containing from 100 to 500 mg of said compound.8. The method according to claim 6 wherein the3-tert.-butyl-4-hydroxyacetanilide is3,5-di-tert.-butyl-4-hydroxyacetanilide.